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An untargeted metabolomics strategy to measure differences in metabolite uptake and excretion by mammalian cell lines

View ORCID ProfileMarina Wright Muelas, View ORCID ProfileIvayla Roberts, View ORCID ProfileFarah Mughal, View ORCID ProfileSteve O’Hagan, Philip J. Day, View ORCID ProfileDouglas B. Kell
doi: https://doi.org/10.1101/2020.06.02.129239
Marina Wright Muelas
1Institute of Integrative Biology, University of Liverpool, UK
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  • For correspondence: m.wright-muelas@liverpool.ac.uk dbk@liv.ac.uk
Ivayla Roberts
1Institute of Integrative Biology, University of Liverpool, UK
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Farah Mughal
1Institute of Integrative Biology, University of Liverpool, UK
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Steve O’Hagan
2School of Chemistry, 131, Princess St, Manchester M1 7DN, UK
3The Manchester Institute of Biotechnology, 131, Princess St, Manchester M1 7DN, UK
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Philip J. Day
3The Manchester Institute of Biotechnology, 131, Princess St, Manchester M1 7DN, UK
4Faculty of Biology, Medicine and Health, The University of Manchester M13 9PL, UK
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Douglas B. Kell
1Institute of Integrative Biology, University of Liverpool, UK
5Novo Nordisk Foundation centre for Biosustainability, Technical University of Denmark, Building 220, Chemitorvet, Kgs Lyngby 2000, Denmark
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  • For correspondence: m.wright-muelas@liverpool.ac.uk dbk@liv.ac.uk
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Abstract

Introduction It is widely but erroneously believed that drugs get into cells by passing through the phospholipid bilayer portion of the plasma and other membranes. Much evidence shows, however, that this is not the case, and that drugs cross biomembranes by hitchhiking on transporters for other natural molecules to which these drugs are structurally similar. Untargeted metabolomics can provide a method for determining the differential uptake of such metabolites.

Objectives Blood serum contains many thousands of molecules and provides a convenient source of biologically relevant metabolites. Our objective was to measure them.

Methods We develop an untargeted LC-MS/MS method to detect a broad range of compounds present in human serum. We apply this to the analysis of the time course of the uptake and secretion of metabolites in serum by several human cell lines, by analysing changes in the serum that represents the extracellular phase (the ‘exometabolome’ or metabolic footprint).

Results Our method measures some 4,000-5,000 metabolic features in both ES+ and ES− modes. We show that the metabolic footprints of different cell lines differ greatly from each other.

Conclusion Our new, 15-minute untargeted metabolome method allows for the robust and convenient measurement of differences in the uptake of serum compounds by cell lines following incubation in serum, and its relation to differences in transporter expression.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted June 02, 2020.
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An untargeted metabolomics strategy to measure differences in metabolite uptake and excretion by mammalian cell lines
Marina Wright Muelas, Ivayla Roberts, Farah Mughal, Steve O’Hagan, Philip J. Day, Douglas B. Kell
bioRxiv 2020.06.02.129239; doi: https://doi.org/10.1101/2020.06.02.129239
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An untargeted metabolomics strategy to measure differences in metabolite uptake and excretion by mammalian cell lines
Marina Wright Muelas, Ivayla Roberts, Farah Mughal, Steve O’Hagan, Philip J. Day, Douglas B. Kell
bioRxiv 2020.06.02.129239; doi: https://doi.org/10.1101/2020.06.02.129239

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